Packing sepharos column.

Packing Sepharose CL-6B
1. Equilibrate all material to the temperature at which the
chromatography will be performed.
2. De-gas the medium slurry.
3. Eliminate air from the column dead spaces by flushing the
end pieces with buffer. Make sure no air has been trapped
under the column net. Close the column outlet with a few
centimeters of buffer remaining in the column.
4. Pour the slurry into the column in one continuous motion.
Pouring the slurry down a glass rod held against the wall
of the column will minimize the introduction of air
bubbles.
5. Immediately fill the remainder of the column with buffer,
mount the column top piece onto the column and connect
the column to a pump.
6. Open the bottom outlet of the column and set the pump to
run at the desired flow rate. This should be at least 133%
of the flow rate to be used during subsequent
chromatographic procedures. However, the maximum flow
rate, see Table 1, is typically employed during packing.
Note: If you have packed at the maximum linear flow
rate, do not exceed 75% of this in subsequent
chromatographic procedures.
7. Maintain the packing flow rate for 3 bed volumes after a
constant bed height is reached.
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